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PMID: 2877932 Published · ppublish English Journal Article Research Support, Non-U.S. Gov't

A straightforward approach to isolate DNA sequences with potential linkage to the retinoblastoma locus.

Human genetics ·Vol. 74 ·No. 3 ·1986-11-00 ·Pages 249-55

Scheffer H, van der Lelie D, Aanstoot GH, Goor N, Nienhaus AJ, van der Hout AH, Pearson PL, Buys CH

Abstract

From a human-Chinese hamster somatic cell hybrid a clone was derived containing chromosome 13 in duplicate as its only human material. This clone was used to construct a human chromosome 13-specific recombinant DNA-library. Overlapping Sau3AI DNA sequences (11.9-17.2 kb) from the cell hybrid were inserted into the lambda phage vector EMBL4. From eleven recombinants having a human insert thirteen putative unique DNA sequences were isolated and cloned into the plasmid vector pBR329. A human-mouse hybrid containing a human chromosome 13 with a deletion of 13q14 and lacking its undeleted homologue was constructed to be used in a selection procedure for DNA sequences belonging to band q14. Three probes originating from two different phages were assigned to 13q14 because they did not hybridise to DNA from this cell hybrid. One of these 13q14 probes detects a low frequency (2/44) MspI restriction fragment length polymorphism. The probes are now being used for screening a cosmid library to find adjacent polymorphic sequences with a RFLP information content suitable for application in the diagnosis of hereditary retinoblastoma.

MeSH Terms
Animals Chromosome Mapping Chromosomes, Human, Pair 13 Cloning, Molecular Cricetinae Cricetulus DNA, Recombinant/isolation & purification Genetic Linkage Genetic Markers Humans Hybrid Cells/analysis Mice Polymorphism, Genetic Polymorphism, Restriction Fragment Length Retinoblastoma/diagnosis,genetics
Chemicals
DNA, Recombinant Genetic Markers
Authors & Affiliations
8 authors, click to expand affiliations / ORCID
Scheffer H
van der Lelie D
Aanstoot G H
Goor N
Nienhaus A J
van der Hout A H
Pearson P L
Buys C H
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Article Info
Journal
Human genetics
Abbr.
Hum Genet
ISSN
0340-6717
DOI
10.1007/BF00282543
Published
1986-11-00
Pages
249-55
Language
English
Region
Germany
NLM ID
7613873
Subset
IM
External Links
PubMed source DOI: 10.1007/BF00282543 View journal details
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